QIAamp 96 DNA QIAcube HT Kit

For automated high-throughput isolation of total DNA from blood, cells, and tissues

Simple and reliable automated processing for cost and time savings

Standardized method suitable for a variety of sample types

High yields even from specialized samples

High-quality DNA that performs well in downstream analyses

Optimized protocols for a range of starting materials

The QIAamp 96 DNA QIAcube HT Kit enables simple, automated purification of total DNA (e.g., genomic, mitochondrial, and pathogen) from blood, cells, and tissue samples. Using proven QIAamp silica-membrane technology in a convenient 96-well format, contaminants and inhibitors are removed to yield high-quality nucleic acids that are ready for downstream analysis. This kit is designed to be used on QIAcube HT System.

Human blood from one donor was treated with sodium citrate, heparin, or EDTA. Genomic DNA was purified from a 50 or 200 µl aliquot of the treated blood manually with the QIAamp 96 DNA Blood Kit or on the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit with the QIAamp 96 DNA QIAcube HT V1.QSP protocol. Purified DNA was eluted in 200 µl. PCR amplification of a 1.2 kb fragment from the single copy Hugl gene was carried out using QIAGEN HotStarTaq DNA Polymerase and 2.5 µl eluate in a total volume of 25 µl. A 1 µl aliquot of each PCR was analyzed on the QIAxcel Advanced with the QIAxcel DNA Screening Kit. M1: pUC18 HaeIII marker; M2: phiX174 HaeIII marker.

High performance in downstream assays.

Genomic DNA was purified from 200 µl human blood. Samples were processed with the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit and the QIAamp 96 DNA QIAcube HT V1.QSP protocol, or with the QIAcube using the QIAamp DNA Blood Mini Kit. Quantitative PCR of IL8 was carried out using the QuantiFast Probe Kit with 5 µL eluate in 25 µl. The CT values obtained show even amplification across the whole 96-well plate, indicating no well-to-well variation in DNA recovery with QIAcube HT. CT values were similar between the different purification platforms.

Purification of genomic DNA from tissues.

Genomic DNA was purified from 5 mg rat lung, kidney, spleen, and liver tissue and 200 µl human blood. Samples were processed with the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit and the QIAamp 96 DNA QIAcube HT V1.QSP protocol or with the QIAcube using the QIAamp Blood Mini Kit (blood) or the QIAamp DNA Mini Kit (tissues). Both platforms showed similar performance in [A] yield and [B] quality of purified DNA for each type of starting material.

Reproducible 96-well DNA purification.

Genomic DNA was purified from 5 mg rat liver with QIAcube HT and the QIAamp 96 DNA QIAcube HT Kit. Quantitative PCR of the prion protein gene was carried out with the QuantiTect SYBR® Green PCR Kit. The resulting CT values show even amplification across the plate, indicating no significant well-to-well variation in DNA recovery.

Automated purification of total DNA from blood samples is quick, easy, and reliable with the QIAamp 96 DNA QIAcube HT Kit. Fresh or frozen whole blood containing common anticoagulants can be processed, and the resulting DNA is of the same high quality as from other QIAGEN purification solutions (see figures Agarose gel of purified genomic DNA and Analysis of genomic DNA from human blood).

QIAcube HT and the dedicated QIAamp 96 DNA QIAcube HT Kit lets users increase sample purification throughput without having to compromise on quality or reliability. The procedure provides high yields of pure DNA that perform well in downstream analyses, similar to other QIAGEN DNA purification solutions (see figures High performance in downstream assays and Purification of genomic DNA from tissues).

Principle

The QIAamp 96 DNA QIAcube HT Kit enables automated purification of total DNA (e.g., genomic, mitochondrial, and pathogen) from a variety of sample sources including fresh or frozen tissues, cells, and blood. Samples treated with common anticoagulants such as citrate, heparin, or EDTA are suitable for use. The procedure yields high-quality DNA that performs well in PCR and other enzymatic reactions.

The QIAamp 96 DNA QIAcube HT Kit combines the selective binding properties of a silica-based membrane with a high-throughput 96-well format, and is designed for fully automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument.

96 samples in approximately 96 minutes
24 samples in approximately 45 minutes

Procedure

The QIAamp 96 DNA QIAcube HT procedure is fast and simple with the QIAcube HT instrument. Samples are first lysed using QIAGEN Proteinase K. Buffering conditions are adjusted to provide optimal DNA binding conditions and the lysate is loaded onto the QIAamp 96 plate. As vacuum is applied, DNA selectively binds to the QIAamp membrane, and contaminants pass through. Remaining contaminants and enzyme inhibitors are removed in three efficient wash steps. Pure DNA is eluted under vacuum in a single step in 200 µl of Buffer AE.

Applications

The QIAamp 96 DNA QIAcube HT Kit provides high-quality DNA, ready to use in various downstream assays, including applications in:

Q Protocol notes for purification of genomic DNA from blood, cells, and soft tissue on the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit; use with the QIAamp 96 DNA QIAcube HT V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

These Q Protocols and installation files can only be used with QIAcube HT Operating software version 4.17 and are not compatible with the new QIAcube HT Prep Manager software.

QIAcube HT Prep Manager Software already includes all standard protocols. Additional protocols and kit configurations can be easily installed. Check for available updates under the "Operating Software" section of the "Product Resources" tab.

Handbook for purification of genomic DNA from blood, cells, and soft tissue on the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit. For use with QIAcube HT Operating Software 4.17; not compatible with QIAcube HT Prep Manager Software.